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Objective To observe the developmental changes of projection and termination of proprioceptive afferent fibers in the mouse spinal cord. Methods Parvalbumin (PV) immunohistochemistry was used to label the proprioceptive afferents. Single and dual immunofluorescence histochemistry were used to examine the growth pattern of proprioceptive afferents and their relationships with motoneurons in the spinal ventral horn (VH). The stained sections were observed under a confocal laserscanning microscope. Results PV-like immunoreactive (LI) proprioceptive fibers first appeared in the dorsal column on embryonic (E) day 14, then entered the gray matter on El5 and reached the intermediate gray matter and VH more obviously on E16. The number and intensity of PV-LI proprioceptive afferent fibers and punctata increased in the VH with age and reached a maximum during earlier postnatal (P) period (P0-P7). After P14, the number and intensity of proprioceptive afferents gradually decreased. The proprioceptive terminals seemed to form close relationship with motoneurons from E17. Conclusion The present study indicates that the somatotopic organization of proprioceptive afferents in the spinal cord is established during the late embryonic and early postnatal stages. These results provide evidence for understanding the development of the reflex movements.
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Objective To observe the relationship between 5-Hydroxytryptamine(5-HT)-like immunoreactive terminals and the vestibulo-parabrachial nucleus projection neurons which may express 5-HT1A receptor in the vestibular nuclear complex(VNC). Methods Retrograded-tract tracing technique combined with double labeling of immunofluorescence histochemical was used,and the stained sections were observed under a confocal laser-scanning microscope. Results Following injection of tetramethylrhodamine(TMR) into the parabrachial nucleus, many retrogradely labeled neurons were observed bilaterally within VNC,but with an ipsilateral predominance.Immunofluorescence histochemical staining showed that many neurons expressed(5-HT1A) receptor-like immunoreactivity and a large number of 5-HT immunostained fibers or terminals were found in the medial,spinal,superior,lateral vestibular nucleus(MVe,SpVe,SuVe,LVe),X nucleus and Y nucleus.Confocal laser-scanning microscopy revealed that some TMR-labeled neurons were 5-HT_1AR immunopositive,and some of the cell bodies or dendrites of TMR/5-HT1AR double-labeled neurons were closely apposed by 5-HT-like immunoreactive terminals.Conclusion The present study suggests that 5-HT may modulate vestibular signals along the VNC-parabrachial nucleus pathway via 5-HT1A receptor.
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Objective To investigate the relationship between the expression of endostatin, VEGFR and chronic altiplano cardiopathy,and the mechanism of the curative effect of rhodiola.on treat altiplano cardiopathy. Methods The rat model with chronic altiplano cardiopathy was created.The experimental rats were divided into four groups: control group;smoking and running stimulated group;altiplano cardiopathy group and rhodiola treated group.The expression levels of endostatin and VEGFR were determined by the methods of immunohistochemistry and semi-quantitative RT-PCR. Results The expression levels of VEGFR in hearts and lungs of the stimulated groups were found higher than that of the control group,while endostatin level was contrary.The expression level of VEGFR in hearts and lungs of the rhodiola treated group was lower than that of the altiplano cardiopathy group,while endostatin was higher.Conclusion The pathogenesis of chronic altiplano cardiopathy seems to be closely associated with epithelial lesion.The curative effect of rhodila to this disease may be exerted by reducing the damage of epithelial cells.
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Objective To investigate the calbindin D\|28K (CB) neurons that receive the visceral nociceptive information in the interstitial nucleus of the spinal trigeminal tract (INV) directly project to the nucleus of the solitary tract (NTS). Methods Triple\|labeled methods of fluorogold (FG) retrograde tracing combined with Fos and CB proteins immunofluorescence histochemistry after formalin stimulating upper alimentary tract. Results Most of FG\|retrograde labeled neurons distributed in the paratrigeminal nucleus (PaV) and the dorsal paramarginal nucleus (PaMd) of INV ipsilateral to the FG injection. About 71^2% of FG\|retrograde labeled neurons contained CB and 31^5% of FG/CB double\|labeled neurons exhibited positive Fos\|immunoreaction in INV.Conclusion The results suggested that a part of neurons containing CB in INV receive the visceral nociceptive information and directly project to NTS. CB neurons might play an important role in transmitting visceral nociceptive information from INV to NTS. [
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Objective To observe the distribution of GABA-containing neurons revealed by GFP expression and the colocalization of the GFP with parvalbumin(PV) in the caudal spinal trigeminal nucleus(Vc),the glutamate decarboxylase 67-green fluorescence protein(GAD67-GFP) knock-in mice were used in the present study. Methods Double-labeled techniques were used by in situ hybridization combined with immunohistochemistry for GFP and double immunofluorescence histochemistry for GFP and NeuN(neuronal nuclei protein,a neuronal marker) or PV.The stained sections were observed under light microscope and a confocal laser-scanning microscope. Results 1.Over 90% of GFP-positive neuronal cell bodies in the laminae Ⅰ and Ⅱ of the Vc showed hybridization signals for GAD67 mRNA,and that almost all neuronal cell bodies with GAD67 mRNA signals were GFP-positive;2.GFP-positive neurons were mainly distributed in the laminae Ⅰ and Ⅱ of the Vc and the vast majority of them were small neurons.The considerable number of GFP-positive processes and somata were most densely observed in the lamina Ⅱ of the Vc.The proportion of GFP-positive neurons in the NeuN-labeled neurons of the Vc was about 19.4% and 24.3% in laminae Ⅰ and Ⅱ,respectively;3.Double-labeled neurons for GFP/PV are mainly found in laminae Ⅰ and Ⅱ of the Vc.The proportion of GFP/PV double-labeled neurons was about 62.4% and 12.8% of total population of PV-and GFP-positive neurons in laminae Ⅰ-Ⅱ of the Vc,respectively.Conclusion GABAergic neurons are mainly distributed in the laminae Ⅰ and Ⅱ of the Vc which related closely to transmission of the nociceptive primary afferent information,and the majority of PV-positive neurons are GABAergic neurons.
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Objective To investigate the expression of endometrial developing-related genes,the endometrium-like structure and cells during spontaneous differentiation of hESCs. Methods (1) Embryoid bodies were cultured in suspension for 14 days,fixed in 10% neutral-buffered formalin and embedded in paraffin.Estrogen receptor(ER) was detected by immunocytochemical staining.(2) hESCs were cultured in a 35 mm plastic dish and differentiated spontaneously for 10 days.The expression of ER and Vimentin/Keratin was detected by double immunofluorescence histochemistry.(3) hESCs were cultured in a 35 mm plastic dish and differentiated for 5 days.The expressions of endometrial developing-related genes including Wnt4,Wnt7a,Wnt5a,Hoxa10,Hoxa11 and ER were detected by RT-PCR. Results (1) The endometrium-like structures were detected in 14-day-old EBs and some were positive for ER staining.(2) Some spontaneously differentiated cells from hESCs for 10 days were positive for ER together with Vimentin/Keratin.(3)Wnt7a,Wnt4,Hoxa10,Hoxa11 and ER were detected by RT-PCR in hESCs which were differentiated spontaneously for 5 days.Conclusion During the spontaneous differentiation of hESCs,some cells are liable to differentiate into endometrial stem cells and endometrium-like cells.However,further identifications of the whole development process are needed to be done in the future.